Original Artcle by Falak Naz, Muhammad Zahoor, Muhammad Ayaz, Muhammad Ashraf, Asif Nawaz, Amal Alotaibi. Presentation prepared by Raktimava Das Sarkar

1. Anti-diabetic potentials of
Sorbaria tomentosa Lindl.
Rehder: Phytochemistry (GC-MS
analysis), α-amylase, α-
glucosidase inhibitory, in vivo
hypoglycaemic, and biochemical
analysis
Presented by
RAKTIMAVA DAS SARKAR
M.Pharm (Pharmaceutics) 2nd Year-3rd
Semester
Roll-19320322015

2. Table 1.Taxonomical
Classification
Kingdom Plantae
Phylum Tracheophyta
Class Magnoliopsida
Order Rosales
Family Rosaceae
Genus Sorbaria
Species
Sorbaria
tomentosa (Lindl.)
Rehd.
Fig 1. Photograph of S. tomentosa

3. Table of
contents
Introduction
Materials and Methods
01
02
Results and Discussion
Conclusion
03
04
Outcomes and Deliveries
05
References
06

4. Journal Details
Journal Name- Open Chemistry
Publisher- De Gruyter Open Access
Impact Factor- 2.3(SCImago as of 2023)
Publishing Since- March 1,2003
H-index- 30
Major Indexing- Web of Science
UGC Approved
SCOPUS
DOAJ
CAS
Publons

5. Publication Details
Article Type- Research
Publish Date- May 6,2023
Authors- Falak Naz, Muhammad
Zahoor, Muhammad Ayaz, Muhammad
Ashraf, Asif Nawaz, Amal Alotaibi

6. 01 Introduction
Diabetes mellitus (DM) is a chronic metabolic disorder of insulin insufficiency due to a
gradual decrease in its production or tissue resistance to its action. The deficiency of insulin
leads to an increase in the concentration of blood glucose level, which gradually
compromises the physiology of vital organs.
The plant of Sorbaria tomentosa Lindl. Rehder belongs to the Rosaceae family. The plant is
a shrub and is 1.5–3 m tall. The leaves are short petiolate and 1–8 cm long and 2–4 cm
wide. The buds are 2 mm long and ovoid, obtuse, and brown.
Project was designed to assess the plant phytochemistry and evaluate it for efficacy
against type 2 DM using in vitro and in vivo approaches.
DM
Type -1insulin
dependent DM
Absolute deficiency
of insulin due to the
destruction of
pancreatic β cells
Type-2 noninsulin-
dependent DM
Gradual decrease
in secretion of
Insulin and tissue
resistance

7. 02
Materials and Methods

8. Extraction Fractionation
A Reddish Black Semisolid mass of methanolic
extract obtained
Filtrate evaporated at 40°C by Rotary
Evaporator
Filtered using muslin cloth, then through filter
paper
Soaked in 80% methanol for 14 days with
occasional shaking
Shade-Dried plant was cut into small pieces and
crushed to a powder
All extracts were collected in separate containers
and solvents removed by Rotary Evaporator
All n-hexane extracts were mixed.
n-hexane layer was separated and then again 500ml
n-hexane was added. Repeated thrice
Stirred vigorously in a separating funnel for some
time to form two layer
500ml of distilled water then 500ml n-hexane was
added

9. GC-MS Analysis Diabetes Induction
To find whether it contains a single compound or
a group of compounds
Interpretation of the mass spectrum was performed
using the National Institute of Standards and
Technology (NIST) database
Identification was done on basis of molecular mass,
molecular structure, and calculated fragments.
GC-MS analyzer (GC Clarius 500 Perkin Elmer),
Elite-1 Column(100% dimethyl polysiloxane) and
carrier gas used was Helium(99.9%)
2 µL methanolic plant extract
The blood glucose
level was recorded after 48 h of administration by a
glucometer.
Animals were kept on a fasting mode for 8–12 h but were
allowed freely to water before bioassay.
Causes
hyperglycemia in mice
Alloxan monohydrate (10%) was administered at a dose
of 160 mg kg−1 body weight
Alloxan monohydrate was obtained
from Sigma Aldrich

10. In-vitro Testing In-vivo Testing
Further Diluted using 10ml DW and absorbance
was checked at 540nm
Reaction was stopped by adding 1 mL of 3,5-
dinitro salicylic acid reagent. Then incubated for 5
min and cooled.
A solution of 1% starch (500 µL) prepared in 0.02
M sodium phosphate buffer was added to each
tube
Incubated for 10 min at 25°C
Test sample and standard drug were added to a
tubecontaining 0.20 mM PBS at pH 6.9 and and
a solution of α-amylase (conc 0.5 mg mL−1)
Group IV: Treatment group received i/p test
compounds (150 and 300 mg kg−1
Group III: Treatment group received
glibenclamide (5 mg kg−1)
Group II: Diabetic group received i/p
alloxan and
Tween 80.
Group I: Control (normal/non-diabetic)
group received
i/p only normal saline.
All animals were grouped into 4 groups
having 6 rats in each group.
0.1 M Na2CO3 soln. was
added to stop the reaction.
Absorption was observed at
405 nm.
3 mM pNPG solution (100 µL)
was added to it and was
incubated for
10 min at 37°C.
pH was adjusted to 6.8. The
solution mixture was mixed
and incubated for 15 min at
37°C.
Various concentrations of
plant samples were mixed
with 50 µL of enzyme soln.

11. Plant Collection and
Verifcation
Aerial plant parts of S.
tomentosa at the
flowering stage were
collected from Murree,
Pakistan, and were
verified by Dr. Ali Hazrat,
a botanical taxonomist at
the Botany Department,
University of Malakand. T
Ethics committee
approval
The animal work of this
project was examined and
approved by the
Departmental Research
Ethics Committee at the
Department of Pharmacy,
University of Malakand,
under reference number
DREC/ST/Diabetes/Pharm-
2020
Experimental animals
Animals used in this study
were rats weighing 145–180 g
with an average weight of
162.5 g. The animals were
placed in an animal house
under standard laboratory
conditions of 25°C temperature
and 12 h light and dark with the
supply of water ad libitum
during the whole experiment
period.
Miscellaneous Information

12. 03
Results and Discussion

13. GC-MS analysis
35 various peaks were observed. The most abundant compounds were oleic acid (20.52%), α-sitosterol (10.44%),
8-octadecenoic acid, methyl ester, (E) (9.11%), trichothec9-en-8-one, 4-(acetyloxy)-12,13-epoxy-3,7,15-trihy droxy-
(3α,4α,7α), (7.57%), phthalic acid, 6-ethyloct-3-yl 2-ethylhexyl ester (7.55%), 9,12,15-octadecatrienoic acid, 2,3-
bis[(trimethylsilyl)oxy]propyl ester, (Z,Z,Z) (7.44%), lupeol (6.02%), α-D-galactopyranose, 6-O-(trimethylsilyl)-cyclic
1,2:3,4-bis (butylboronate) (5.15%), and α-sitosterol trimethylsilyl ether (5.06%)
Figure 2. GC-MS Spectra
Figure 3. Identified compounds

14. In-vitro Study Inhibition of α-amylase and α- glucosidase
The most potent extract was chloroform, which
exhibited an inhibition of 84.4 ± 1.12% of the enzyme
at a concentration of 1,000 µg 𝑚𝑙−1
.
IC50 for the crude methanolic extract was 530 µg𝑚𝑙−1
thus it exhibited concentration-dependent moderate
enzyme inhibitory potentials.
Figure 5. Inhibitory potentials of α-glucosidase of the crude sample
from Sorbaria tomentosa. Statistical significance was set as p < 0.05.
Figure 4. Inhibitory potentials of α-amylase in various solvent extracts
and standard drugs. Statistical significance was set as p < 0.05.

15. In-vivo Studies
1. ACUTE TOXICITY STUDY
No morbidity or mortality was observed.
No aberrant behaviour was observed during the acute toxicity studies at the tested doses.
2. ANTI-HYPERGLYCEMIC EFFECT
Diabetic animals treated with the
standard drug exhibited a gradual
decrease in blood glucose during
therapy.
The treatment of diabetic animals
with 150 mg kg−1 dose of crude
methanolic extract also showed a
steady decrease in blood glucose
levels. Furthermore, the crude
methanolic extract at a dose of 300
mg kg−1 exhibited lowered blood
glucose levels.
Figure 6. Results of in vivo anti-diabetic potentials of S. tomentosa.

16. In-vivo Studies
3. CHANGES IN THE WEIGHT OF VITAL ORGANS
Overall, no major changes were recorded during therapy in the weight of these organs
Table 2. Effects of the crude extract treatment on the weights (g) of vital organs in fasting rats

17. 04 CONCLUSION
 The results of the phytochemical analysis of the study revealed that plants
contain various metabolites, which might be responsible for the anti-diabetic
potentials of the plant.
 Various extracts exhibited concentration-dependent inhibition of key
enzymes implicated in postprandial hyperglycemia and type-2 diabetes.
 The crude methanolic extract showed considerable in vivo efficacy against
alloxan-induced diabetes in rats.
 Further studies include activity-guided isolation of bioactive metabolites, and
molecular studies are required for further results.

18. 05 OUTCOMES AND DELIVERIES
 The anti-diabetic activity has been proved and the compounds have been
identified as a whole. But which compound is the prime phyto-component
that shows this anti-diabetic property is unknown.
 In vivo studies needed to be performed with other solvents. Testing with
Methanolic extract was performed, Chloroform, Ethyl Acetate and n-Hexane
was not performed.
 Acute toxicity study data is not shown.
 HPTLC and Electrophoresis might be helpful in finding the active
metabolite.
 Molecular Docking can be a resourceful tool in finding the active site.

19. 06 REFERENCES
1. Iftikhar H, Ahmed D, Qamar MT. Study of phytochemicals of Melilotus indicus and alpha‐amylase
and lipase inhibitory activities of its methanolic extract and fractions in different solvents.
ChemistrySelect. 2019;4(26):7679–85.
2. Chen L, Kang Y-H. In vitro inhibitory effect of oriental melon (Cucumis melo L. var. makuwa Makino)
seed on key enzyme linked to type 2 diabetes: Assessment of anti-diabetic potential of functional
food. J Funct Foods. 2013;5(2):981–6
3. Szkudelski T. The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas.
Physiol Res. 2001;50(6):537–46. [29] Ngugi MP, Kimuni N, Ngeranwa N, Orinda O, Njagi M, Maina
D, et al. Antidiabetic and safety of Lantana rhodesiensis in alloxan induced diabetic rats. J Dev Soc.
2015;4(1):129–38.
4. Lakshmi V, Mahdi AA, Ahmad MK, Agarwal SK, Srivastava AK. Antidiabetic activity of lupeol and
lupeol esters in streptozotocin-induced diabetic rats. Bangladesh Pharm J. 2014;17(2):138–46.
5. Queen ZE, Rao M, Anthony J, Prabhu K, Johnson W, Balasubramanian BS, et al. The GC MS
study of one ayurvedic preparation amrithamehari Churnam. Int J Pharm Sci Rev Res.
2016;39(2):169–72
6. Franz MJ, Boucher JL, Rutten-Ramos S, VanWormer JJ. Lifestyle weight-loss intervention
outcomes in overweight and obese adults with type 2 diabetes: a systematic review and
metaanalysis of randomized clinical trials. J Acad Nutr Dietetics. 2015;115(9):1447–63.
7. Salehi B, Ata A, V. Anil Kumar N, Sharopov F, RamirezAlarcon K, Ruiz-Ortega A, et al. Antidiabetic
potential of medicinal plants and their active components. Biomolecules. 2019;9(10):551

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